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Objective:To discuss the expression of programmed cell death-ligand 1(PD-L1)in the oral squamous cell carcinoma(OSCC)cells and its effect on biological behavior of the OSCC CAL27 cells,and to clarify the possible mechanism.Methods:Western blotting method was used to detect the expression levels of PD-L1 protein in the oral epithelial HOK cells and OSCC CAL27,TCA8113,and SCC15 cells;immunofluorescence staining method was used to detect the expression and localization of PD-L1 protein in the CAL27 cells.The CAL27 cells were divided into control group(transfected with si-NC)and si-PD-L1 group(transfected with si-PD-L1).Western blotting method was used to detect the interference efficiency of the cells in two groups;CCK-8 assay was used to detect the proliferative activities of the cells in two groups at different time points;plate clone formation assay was used to detect the numbers of clone formation of the cells in two groups;cell scratch healing assay was used to detect the scratch healing rates of the cells in two groups;Transwell chamber assay was used to detect the numbers of migration and invasion cells in two groups.Results:The expression level of PD-L1 protein in the OSCC cells was higher than that in the HOK cells(P<0.05 or P<0.01);PD-L1 expressed in the cytoplasm and nucleus of the CAL27 cells.The CCK-8 assay and plate clone formation assay results showed that compared with control group,the proliferative activities of the CAL27 cells in si-PD-L1 group at different time points were significantly decreased(P<0.05 or P<0.01),and the numbers of clone formation were significantly decreased(P<0.01).The cell scratch healing assay results showed that compared with control group,the scratch healing rates of the cells in si-PD-L1 group were significantly decreased(P<0.05 or P<0.01).The Transwell chamber assay results showed that compared with control group,the numbers of migration and invasion cells in si-PD-L1 group were significantly decreased(P<0.01).Conclusion:The expression of PD-L1 in the OSCC cells is higher than that in normal oral epithelial cells,and knocking down PD-L1 expression can inhibit the proliferation,clone formation,migration and invasion capabilities of the OSCC cells.
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【Objective】 To establish the optimal treatment model of Bletilla striata polysaccharide (BSP) on dextran sodium sulfate (DSS)-induced ulcerative colitis (UC) in mice. 【Methods】 We randomly divided 48 male C57BL/6 mice into normal control group, DSS model group (25 g/L DSS), BSP low-, medium- and high-dose groups (25 g/L DSS + 95, 190, 380 mg/kg BSP), and salazosulfapyridine (SASP) (25 g/L DSS + 320 mg/kg SASP, positive control) group. Mice in the normal control group drank distilled water freely, while the other groups were given 25 g/L DSS solution to drink freely for 7 days. From the second day, the low-, medium- and high-dose BSP groups and SASP (positive control)group were administered by gavage according to body mass. The normal control group and DSS model group were given the same amount of normal saline once a day for 7 consecutive days. The mice’s blood pressure was recorded every day. Mental state, body mass, stool characteristics and bloody stool were used to calculate the mice’s disease activity index (DAI). The mice were killed on the 9th day, and their colonic tissues were taken for hematoxylin eosin (HE) staining and histopathological scoring. The expression of tight junction protein Claudin-1 in colonic tissues was detected by Western blotting. 【Results】 Compared with the normal control group, the DSS model group had obvious clinical manifestations, histopathological changes and reduced body weight, increased histopathological score and DAI score (P<0.05), and decreased expression of tight junction protein Claudin-1 in colon tissue (P<0.05). Compared with those in DSS model group, the clinical manifestations of UC and colonic mucosal injury in low-, medium- and high-dose BSP groups were improved in varying degrees. The high-dose (380 mg/kg) BSP group had the best effect. The degree of body weight reduction, histopathological score and DAI score in this group were significantly lower than those in DSS model group (P<0.05), whereas the expression of Claudin-1 increased significantly (P<0.05). 【Conclusion】 When BSP was administered by gavage at 380 mg/kg, the therapeutic effect on UC mice induced by 25 g/L DSS was the best. This model can be used as an effective one for further studies on Striata Bletilla polysaccharide in UC mice.
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Objective To clarify the relationship between CT120,a novel human plasma membrane-associated gene,and the proliferation of lung adenocarcinoma cell line A549.Methods Expression vector(pcDNA3.1)containing antisense oligonucleotides of CT120 was constructed and transfected into the adenocarcinoma cell line A549.RT-PCR and Western blot detected the expression of CT120.Meantime,flow cytometry and soft agarose colony formation were applied for cell proliferation,and P53,CyclinD1 and CDK4 were detected by RT-PCR.ResultspcDNA3.1 containing antisense oligonucleotides of CT120 was successfully constructed and inhibited the expression of CT120 effectively by RT-PCR and Western blot.The expression of P53 was up-regulated and the expression of CyclinD1 and CDK4 were down-regulated.Conclusion The down-regulation of CT120 expression by antisense oligonucleotides technique may be a potential drug target for treatment of lung cancer.
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AIM:To investigate the mechanism of 9-cis-RA inhibiting the growth of lung adenocarcinoma cells, and we detect the expressional changes of cyclinD1 and cdk4 in lung adenocarcinoma cells PG, A_(549), SPC-A_1 before and after being treated with 9-cis-retinoic acid(9-cis-RA). METHODS: RT-PCR was used to analyse the transcriptional changes of cyclinD1 and cdk4 in PG, A_(549) and SPC-A_1. RESULTS:9-cis-RA decreased the transcription of cyclinD1 in PG and A_(549)(P
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To study effects of 9 cis retinoic acid (9 cisRA) on biological characteristics of lung squamous cancer cells, cell growth, cell differentiation and apoptotic indexes were observed in 9 cisRA treated L 78 and A 2 lung squamous cancer cells with flow cytometry. The results showed that 9 cisRA exerted marked inhibitory effect on the growth of two lung squamous cancer cell lines, 9 cisRA also had significant inducing effect on the differentiation of L 78 and A 2 cell lines, whereas the percentages of apoptosis of two cell lines in the treatment group were significantly higher than the control group. We conclude that 9 cisRA could inhibit growth inhibition, induce cell differentiation and apoptosis of L 78 and A 2 lung squamous cancer cells